After the intravenous transplantation of MSCs, a significant population of cells accumulates in the lung, which they alongside immunomodulatory effect could protect alveolar epithelial cells, reclaim the pulmonary microenvironment, prevent pulmonary fibrosis, and cure lung dysfunction. The fact that the transplantation of MSCs improved the outcome of COVID-2019 patients may be due to regulating inflammatory response and promoting tissue repair and regeneration. This is a preliminary report of our study in Iran.

1. Mesenchymal Stem Cell Treatment
for Pneumonia Patients Infected
With COVID-19
Preliminary Report
Amir Abbas Hedayati-Asl
Ped. Hematologist Oncologist /BMT
Tehran University of Medical Sciences, HORC-SCT
Royan Institute
Cancer Personalized Medicine Program
Stem Cell Biology and Technology Department

2. SARS-CoV-2
• Infection caused clusters of severe respiratory illness similar to
severe acute respiratory syndrome coronavirus and was
associated with ICU admission and high mortality.
• There is no confirmed antivirus therapy for people infected
SARS-CoV-2, most of them should receive supportive care to
help relieve symptoms.
• For severe cases, treatment should include care to support
vital organ functions. This clinical trial is to inspect the safety
and efficiency of Mesenchymal Stem Cells (MSCs) therapy for
pneumonia patients infected with SARS-CoV-2.

3. SARS-CoV-2
• There is currently no vaccine and no specific antiviral
treatment recommended for SARS-CoV-2 infection.
• About 20% of the patients were severe and some
died of respiratory failure or multiple organ failure.
• Therefore, it is urgent to find a safe and effective
therapeutic approach to pneumonia patients
infected with SARS-CoV-2.

4. Mesenchymal Stem Cells
• In the last year, the promising
features of mesenchymal stem
cells (MSCs), including their
regenerative properties and
ability to differentiate into
diverse cell lineages, have
generated great interest
among researchers whose
work has offered intriguing
perspectives on cell-based
therapies for various diseases.

5. MSC Effects

6. Mesenchymal Stem Cells
• These findings seem to
highlight that the
beneficial effect of MSC-
based treatment could be
principally due by the
immunomodulation and
regenerative potential of
these cells.

7. Immunomodulatory
• The MSCs exert their immune suppressive
potential by cell to cell contact and by
secretion of immune regulatory molecules.
MSCs display broad immunomodulatory
properties, MSCs can inhibit the proliferation
and function of T cells, Natural killer T (NKT)
cells, T regulatory cells， B cells and
dendritic cells (DCs). However, MSCs
secrete IL-6 and preserveneutrophils viability
by inhibiting apoptosis. Several soluble
factors have been shown to play a major role
in theimmunosuppressive effects of MSCs,
including prostaglandin
E2 (PGE2), transforming growth factor (TGF)-
β1, indoleamine 2,3-dioxygenase (IDO), nitric
oxide, hepatocyte growth
factor(HGF), interleukin (IL)-6 and IL-10.

8. UC-MSC
• The investigators found that infusions of UC-MSC
significantly improved liver function in decompensated
liver cirrhosis and primary biliary cirrhosis (PBC) patients,
increased the survival rate in acute-on-chronic liver
failure (ACLF) patients .
• MSCs could significantly reduce the pathological changes
of lung and inhibit the cell-mediated immune
inflammatory response induced by influenza virus in
animal model .

13. * 31 Study in Clinicaltrial.gov
* Only 7 trials Recruiting

14. Purpose of Study
• Investigate safety and efficiency of MSCs in treating pneumonia
patients infected with SARS-CoV-2.
• This trial recruit 6 patients.
• Patients received i.v. transfusion one round (3 times) of 7.5*10⁷ cells
of MSCs as the treated group, all of them received the conventional
treatment.
• In addition, the equal 6 patients received conventional treatment
were used as control.
• The clinical symptoms, pulmonary imaging, side effects, 28-days
mortality, immunological characteristics (immune cells,
inflammatory factors, etc.) be evaluated during the 100 days follow
up.

15. Study Design
• Study Type : Interventional (Clinical Trial)
• Estimated Enrollment : 12
• Intervention Model: Parallel
• Assignment Masking : None (Open Label)
• Primary Purpose: Treatment
• Official Title: Safety and Efficiency of
Mesenchymal Stem Cell in Treating
Pneumonia Patients Infected With COVID-19

16. Clinical classification of the COVID-19

17. Study Design
• Experimental: MSCs
Treatment Group
Conventional treatment
plus MSCs
• Participants receive
conventional treatment
plus 3 times of
MSCs(7.5*10⁷ MSCs
intravenously at Day 0,
Day 3, Day 6).

18. Intervention/treatment
• Biological: MSCs
• 3 times of
MSCs(7.5*10⁷ MSCs
intravenously at Day 0,
Day 3, Day 6).

19. Outcome Measures
Primary Outcome Measures :
Size of lesion area by chest radiograph or CT
Time Frame: At Baseline , Day 3, Day 6, Day 10, Day 14
Evaluation of Pneumonia Improvement
Side effects in the MSCs treatment group:
Time Frame: At Baseline , Day 3, Day 6, Day 10, Day 14, Day 21, Day 28, Day 100
Number of participants with treatment-related adverse events as assessed

20. Secondary Outcome Measures
• Improvement of Clinical symptoms including duration of fever and respiratory
Time Frame: At Baseline , Day 3, Day 6, Day 10, Day 14, Day 21, Day 28
Evaluation of Pneumonia Improvement
• Time of nucleic acid turning negative
Time Frame: At Baseline , Day 3, Day 6, Day 10, Day 14, Day 21, Day 28, Day 90
Marker for COVID-19
• Rate of mortality within 28-days
Time Frame: Day 28
Marker for efficacy of treatment
• CD4+ and CD8+ T celll count
Time Frame: At Baseline , Day 3, Day 6, Day 10, Day 14, Day 21, Day 28, Day 90
Marker of Immunological function

21. • Alanine aminotransferase
Time Frame: At Baseline , Day 3, Day 6, Day 10, Day 14, Day 21, Day 28, Day 90
Markers of organ function
• C-reactive protein
Time Frame: At Baseline , Day 3, Day 6, Day 10, Day 14, Day 21, Day 28, Day 90
Markers of Infection
• Creatine kinase
Time Frame: At Baseline , Day 3, Day 6, Day 10, Day 14, Day 21, Day 28, Day 90
Markers of organ function

22. Eligible for Study & Criteria
Inclusion Criteria:
• Male or female, age 18 years -70 years old
• Confirmed COVID-19 by RT-PCR from any diagnostic sampling source
• Pneumonia that is judged by chest radiograph or computed tomography
Exclusion Criteria:
• Patients with malignant tumor, other serious systemic diseases and psychosis;
• Patients who are participating in other clinical trials
• Inability to provide informed consent or to comply with test requirements.
• Co-Infection of HIV, tuberculosis, influenza virus, adenovirus and other respiratory
infection virus

23. Ethics Certificate

25. Schematic of the Isolation of Cells from Human Umbilical Cord,
Cord-placenta Junction, and Fetal Placenta
Step 1. Manually dissect and separate the
cord/placenta sample into three discrete regions:
umbilical cord (UC), cord-placenta junction (CPJ),
and fetal placenta (FP). Separate the two distinct
zones of the UC into cord lining (CL) and
Wharton's Jelly (WJ) using scissors and forceps.
Step 2. Cut each of the dissected tissues separately
into 1- to 2-mm pieces using scissors and partially
digest the pieces.
Step 3. Culture the tissue pieces.
Step 4. Isolate the cells from the explants by trypsin
treatment. Subculture and characterize the
isolated cells.
Step 5. Isolated and characterized cells can be
amplified and used for various applications.

26. Harvest method Summary

27. Dissection and Culture of Explants from Human Umbilical Cord,
Cord-placenta Junction, and Fetal Placenta.
• three different anatomical
regions of the cord/placenta
sample:
• UC (split into the CL and WJ), CPJ,
and FP, as well as the associated
arteries and veins. CL, WJ, CPJ,
and FP were separated by manual
dissection to isolate cells from
the explants.
• Each of the dissected regions was
separately cut into small
fragments and cultured in 75-
cm2 plates using 9 mL of CM.

28. Morphology of cultures 48 hr after isolation and
removing the tissue debris
• (A) The appearance of the culture
supernatant can vary substantially between
placenta donors before washing off the
debris.
• Example cultures 1 and 2 demonstrate this
variation. These two isolations were
performed simultaneously, but from two
different placentas. Once washed cultures
will be clear of RBCs and tissue debris as
shown in example 3. The subsequent
expansion results are generally consistent.
• (B) Following the 48 hr media exchange,
only a few cells will be attached to the flask.

29. Study Results

31. Patients & Outcome
• Patients: 6 ( 4 Male, 2 Female)
• Age: Mean 45yr (25- 63 yr)
• Survive: 5 pts alive, 1 pt death( PTE)
• Disease: DM 4, Htn 4, IHD 3, AKI 2, Multi Organ failure 1
• MSC, microbial culture: Negative

32. Patients & Outcome
• Before the MSC transplantation, the patients had
symptoms of high fever (38.5℃ ± 0.5℃), weakness,
shortness of breath, and low oxygen saturation.
• 2-4 days after transplantation, the symptoms
disappeared in 4 patients, the oxygen saturations
rose to ≥ 95% at rest, without or with oxygen uptake
(5 liters per minute).

33. Patients & Outcome
• No acute infusion-related or allergic reactions were
observed within two hours after transplantation.
• Similarly, no delayed hypersensitivity or secondary
infections were detected after treatment.

34. Secondary Outcome Measures
After treatment
The peripheral lymphocytes
were increased,
The C-reactive protein
decreased Significant
Response(p<0.05)
ALT were decreased.
Sever & Critical
LDH↑
Alb↓
PT ↑
CRP↑
Lymph↓
ALT ↑
Cr ↑

35. CD4 & CD8 Before /After
After treatment, the peripheral lymphocytes were increased, CD4+ cells and
CD8+ cells increased 3-6 days in 4 pt’s.

36. Radiology Finding
• The primary findings on CT in pt’s reported:
• ground-glass opacities (GGO): bilateral,
subpleural, peripheral
• crazy paving appearance (GGOs and inter-
/intra-lobular septal thickening)
• air space consolidation
• bronchovascular thickening in the lesion
• traction bronchiectasis
• The ground-glass and/or consolidative
opacities were bilateral, peripheral, and
basal in distribution .
• CT Scan Score:
• Before treatment:
Mean 20 ( 16- 25) grade 4
• After Treatment :
Mean 5 ( 0-12) grade 1
Significant Response(p<0.05)

37. Case 1: two axial lung ct scan without contrast in a 51 year old male with covid 19 pneumonitis: before
treatment (A) bilateral and multifocal patchy consolidation, after treatment(B) dramatic response to
treatment and complete resolution of opacities

38. Case 2: two axial lung ct scan without contrast in a 63 year old female with covid 19 pneumonitis:
before treatment (A) bilateral and multifocal patchy consolidation , GGO , with bilateral pleural
effusion, after treatment(B) dramatic response to treatment and nearly complete resolution of
opacities
A B

39. Case 3 : two axial lung ct scan without contrast in a 50 year old male with covid 19 pneumonitis: before
treatment (A) bilateral and multifocal patchy consolidation with bilateral pleural effusion, after
treatment(B) dramatic response to treatment and nearly complete resolution of opacities

40. Case 4: two axial lung ct scan without contrast in a 25 year old male with covid 19 pneumonitis: before
treatment (A) large area of consolidative opacity at right upper lobe and multifocal patchy GGO &
consolidation at left upper lobe, after treatment(B) dramatic response to treatment and complete
resolution of opacities

41. Conclusion:
• MSCs could cure or significantly improve the
functional outcomes of six patients without
observed adverse effects.
• The pulmonary function and symptoms of these
four patients were significantly improved in 3
days after MSC transplantation.
• Thus, the intravenous transplantation of MSCs
was safe and effective for treatment in patients
with COVID-19 pneumonia, especially for the
patients in critically severe condition.

42. Our Team:
• Prof. Ghavamzadeh
• Prof. Abolghasemi
• Dr. Mousavi
• Dr. Nikbakht
• Dr. Bahramifar
• Dr. Imani
• Dr. Jafari
• M. Haghighi
• A. Ghashghaei
• Dr. Hedayati-Asl

43. Acknowledgement
This work was supported by the :
*HORCSCT
*BMSU
Conflicts of interest
We have no conflicts of interest.

44. Thank You
Our Best Hope for Fighting Coronavirus